1997年 · 第110卷第02期

OBJECTIVE To develop a multiprimer-polymerase chain reaction (multiprimer-PCR) method for detecting immunoglobulin heavy chain (IgH) and T-cell receptor (TCR) rearranged genes in the same amplification. METHODS Multiprimer-PCR protocol, a mixture of four different primers used in the same reaction detected IgH CDR-Ⅲ and TCR VγI-Jγ Ligenetic rearrangements in 40 acute lymphoblastic leukemia (ALL) patients. RESULTS Thirteen cases were found to have IgH CDR-Ⅲ and TCR VγI-Jγ genetic rearrangements. Ten cases were found to have TCR VγI-Jγ genetic rearrangements. Thirteen cases were found to have IgH CDR-Ⅲ genetic rearrangements. The amplification result of multiprimer-PCR was the same as separate amplification. The sensitivity of multiprimer-PCR was 10-4-10-5 DNA level, which was the same as separate PCR amplification. All positive cases with IgH rearranged gene were confirmed by Southern blot. CONCLUSIONS Multiprimer-PCR could screen two distinct rearranged genes in a single amplification, and is more suitable for screening of specimens collected at diagnosis.

OBJECTIVE To determine whether mRNA for endothelin-1 was present and changed in the lungs of rats exposed to chronic hypoxia. METHODS cRNA probes for rat ET-1 and c-fos were labelled by in vitro transcription with digoxigenin-UTP for in situ hybridization. Endothelin-1 like immunoreactivity (ET-1-LI) in plasma as well as in lung homogenate was measured by radioimmunoassay. RESULTS The vascular endothelial cells of pulmonary arteries and the bronchiolar epithelial cells were strongly positive for the ET-1 probe. The perivascular and peribronchiolar smooth muscle were positive for the c-fos probe, primarily. There was little expression of ET-1 and c-fos in the rat lungs in controls. There was significant increase of ET-1-LI levels in rats exposed to hypoxia for 3 weeks, the concentration ET-1-LI in venous samples was 3.85±1.52 ng/L (P<0.05 as compared with the control groups), in arterial sample was 4.72±1.66 ng/L (P<0.05) and in lung was 2.06±0.68 ng/g wet lung weight (P<0.05). CONCLUSIONS Chronic hypoxia elevated the mean pulmonary arterial pressure, caused vessel remodelling and the right ventricular hypertrophy. These changes were accompanied by an increase of ET-1 in plasma and lung homogenate. The expression and production of ET-1 were localized to endothelium and airway epithelium in the lungs.

OBJECTIVE To evaluate the diagnosis and aggressive surgical approach for brainstem tumors. METHODS A series of 24 adult patients with brainstem tumors were involved. Magnetic resonance (MR) imaging was performed on all patients. Surgical technique, potential risk and management of complications were described. RESULTS Diagnosis was confirmed by MR imaging and histological examination in all the patients. Tumors were totally or subtotally removed in twenty-two patients. Signs and symptoms were improved in 19 patients on discharge. CONCLUSIONS Our study shows that MR imaging has an important value for localizing tumors, identifying their nature and selecting appropriate surgical approaches. Most brainstem tumors, except for high-grade gliomas and small ventral tumors, are amenable to an aggressive surgical approach. The favorable results depend upon the appropriate surgical approach, microsurgical technique and intensive care after operation. Total removal can provide both high-quality survival and favorable long-term prognosis.

OBJECTIVE To explore a more effective immunotherapy for cancer by studying tumor-infiltrating lymphocytes (TIL) of patients with hepatocarcinoma. METHODS TIL isolated from freshly resected tumors of 12 patients with hepatocarcinomas were activated, expanded and cultured in vitro by incubation with interleukin-2, and then infused to the patients. The TIL cytotoxicity and phenotype were determined by MTT and immunohistochemical ABC methods. RESULTS The average weight and number of cells of the tumor tissues for culture of TIL were 4.8 g and 5.8×10 7 respectively, and the median expansion time was 31.8 days in vitro. The maximal expansion achieved 1000 fold, and the mean amplification rate was 198.5 fold. Lymphokine-activated killer cell culture supernatant could stimulate TIL growth. Ten of 12 patients received immunotherapy of TIL. The number of cells infused reached 4×108-1.1×1010. In 3 cases of our series, TIL infusions were done through a drug pump with a catheter embedded in the hepatic artery. Eight patients whose primary hepatic carcinomas were resected completely were followed up for 16 months with only one case (12.5%) having recurrence. The recurrence rates of them at 6 and 12 months after tumor resection were remarkably lower than those of the patients who did not receive TIL infusion (19.4% and 41.6%, respectively). One patient whose tumor was not able to be removed received TIL infusion twice and local alcohol injection several times. He has been alive for 9 months. CONCLUSIONS TIL isolated from the tumors of patients with hepatocarcinoma can be activated and expanded in vitro. It demonstrated remarkably cytotoxic activity against tumor targets. TIL infusion is an effective immunotherapy for patients with hepatocarcinoma in reducing recurrence after operation.

OBJECTIVE To examine whether there is generation of oxygen free radicals (OFR) and lipid peroxidation of cell membrane after volume replacement for burn shock, and to study the relationship between OFR injury and enterogenous endotoxemia. METHODS Forty-seven burn patients were involved in this study. Among them, 18 had delayed fluid resuscitation (DR) and the others had early fluid resuscitation (ER) within 6 hours postburn. Sixty-six gnotobiotic rats were used in a collaborating experiment as burn models. They were divided into 4 groups: sham injury (n=6), early resuscitation (n=24), late resuscitation (n=24) and vitamins E and C treatment group (n=12). All the rats, except those in the sham injury group, were inflicted with 40% total body surface area (TBSA) third-degree burns. OFR was determined in the blood of patients with electron spin resonance (ESR). S/W ratio and tau c values of patients' erythrocytes were measured with ESR spectrometer. Blood superoxide dismutase (SOD) and glutathione peroxidase (GSHPx) activities, malondialdehyde contents and plasma endotoxin levels were assayed. Rats were sacrificed at the 12th, 24th, 48th and 72nd hour after injury. Plasma endotoxin levels, mucosal SOD, GSHPx and malondialdehyde (MDA), as well as diamine oxidase activity of ileum were determined. Cultures of mesenteric lymph nodes (MLN), liver, spleen, heart, lung, kidney and blood were done. RESULTS A significant increase in blood OFR contents and plasma MDA, and a significant decline in blood SOD and GSHPx were found after resuscitation in DR group as compared with those in ER group. Both strong to weak spectra component (S/W) ratio and tau c value were higher in DR group in contrast with those in ER group. Higher elevation in plasma endotoxin level in DR group was seen. In DR group, plasma MDA content was correlated with S/W ratio, tau c value and plasma endotoxin level. In rats, the level of mucosal MDA, plasma endotoxin and incidence of bacterial translocation (BT) were significantly higher. Mucosal SOD, GSHPx and diamine oxidase (DAO) activity were significantly lower in DR group as compared with those in ER group. In DR group, mucosal MDA content was negatively correlated with mucosal DAO activity, while the latter was negatively correlated with BT. After treatment with vitamins E and C, mucosal MDA content decreased, plasma endotoxin and BT significantly declined and mucosal DAO heightened. CONCLUSIONS Tissue reperfusion might induce the production of OFR, resulting in lipid peroxidation injury, especially to intestinal mucosa, and resulting in disruption of mucosal barrier function followed by endotoxemia and BT.

OBJECTIVE To compare the effect of vascularized and nonvascularized fibula grafting performed for the reconstruction of bone defects after the resection of bone tumors. METHODS Vascularized fibula grafting was performed in 18 patients with malignant or aggressive benign bone tumor of the extremities. Non-vascularized fibula grafting was performed in 15 patients with giant cell tumor of the radius. All the 18 patients were followed up with radiography, single photon emission computed tomography (SPECT), colored Doppler's sonography (CDS) examinations and functional observation for 2 to 14 years. RESULTS Vascularized fibula grafts with bone defects of 9 to 26 cm showed good blood supply in emission computerized tomography (ECT) and radiography examinations, and bone union was achieved (Table 1). In non-vascularized fibula grafting cases (Table 2), bone union was not achieved. CONCLUSIONS The advantages of vascularized grafting are the feasibility of one-stage reconstruction of various tissues and the growth of grafted fibula. It is necessary to adopt vascularized fibula grafting to reconstruct large bone defect resulting from the resection of extensive bone tumors, such as giant cell tumor (GCT), fibrous dysplasia of bone and malignant bone tumors.

OBJECTIVE To examine the difference in the light sensitivity in the central visual field between the normal subjects and patients with or without the early visual field damages and propose the criteria to identify the possible early disturbances in the visual field. METHODS Light sensitivities at the 76 locations in the central field were measured in 108 normal eyes of 75 normal volunteers aged from 20 to 78 years, and 82 eyes of 53 patients with early open-angle or closure-angle glaucoma on the Humphrey Field Analyzer (Model 610), using the Central-30-2 program. RESULTS No sex and eye differences were found. The significant differences in the light sensitivities existed between the age of 20-45 years and the age of 46-78 years in the normal subjects. The lower limit of 80% normal range for each location in the central field of the two age groups was calculated. The light sensitivity decreased by 2-3 dB at every 10 degree from the eccentricity. The light sensitivity in the inferior field was 1-2 dB higher than that in the superior field. There was no statistical difference between the nasal and the temporal field. For the patients with the field defects, the light sensitivity of many locations decreased by 4-8 dB, and by 1-2 dB in the patients without the field defects as compared with the normal controls. CONCLUSIONS Based on comparisons with the normal controls and the lower limit of 80% normal range, we proposed the following criteria for classification of the damages in the glaucoma visual field. Grade I: light sensitivity reduces by 1-3 dB at a cluster of locations in the field; Grade II: light sensitivity reduces by 4-6 dB at a cluster of locations in the field; Grade II: light sensitivity reduces by more than 6 dB at a cluster of locations in the field.

OBJECTIVE To further carry out a quantitative immunohistochemical study on neuropeptides (NPs) participation in perennial allergic rhinitis (PAR) in nasal mucosa. METHODS By using light microscopy (routine hematoxylin and eosin staining, Wright's staining and 1% toluidine blue staining) and scanning as well as transmission electron microscopy, an exhaustive immunohistochemical and morphometric study was made on the changes of substance P (SP), neuropeptide A (NKA) and neuropeptide B (NKB) in the nasal mucosa of a toluene-2,4-iso-cyanate (TDI)-induced PAR model of rat. RESULTS The results showed that the densities of all three peptidergic terminals were significantly increased (P<0.01) in experimental group (115.72±28.43 for SP, 39.23±10.34 for NKA and 37.24±11.22 for NKB, respectively) as compared with those in control group (49.65±11.23 for SP, 12.44±6.21 for NKA and 13.31±7.60 for NKB, respectively). Increased staining, thickening of peptidergic terminals and enlargement of varicosities were found. Mast cell infiltration in nasal mucosa was in parallel with increase of peptidergic terminals. CONCLUSIONS All results support the hypothesis that NP is a newly-recognized but potentially important factor in the pathogenesis of PAR.

OBJECTIVE To examine the role of otoacoustic emissions and auditory efferent system function in evaluating auditory functional status in both normal and diseased conditions. METHODS Spontaneous and transient evoked otoacoustic emissions and efferent function in terms of contralateral white noise induced transient evoked otoacoustic emission suppressions were tested and cross compared in 32 normal ears, 24 ears with cochlear hearing loss and 16 ears with retrocochlear hearing loss. RESULTS Transient evoked otoacoustic emission amplitude and contralateral suppression, spontaneous otoacoustic emission incidence, peaks and frequency range were significantly reduced in cochlear hearing loss while in retrocochlear hearing loss transient evoked otoacoustic emission and spontaneous otoacoustic emission levels were significantly higher than in cochlear hearing loss group and showed no suppression. CONCLUSIONS Contralateral acoustic stimulation induced transient evoked otoacoustic emission suppression as an index of efferent function is clinically feasible. Combined otoacoustic emission and efferent tests are of great significance in evaluating cochlea status and auditory central mechanisms. Auditory efferent function is weakened in cochlear lesions and severely damaged in retrocochlear lesions.

OBJECTIVE To investigate the expression of nm23-H1 in epithelial ovarian cancer. METHODS A total of 55 specimens from patients with epithelial ovarian cancers, 15 patients with serous cancers, 7 with mucinous cancers, 17 with endometrioid cancers and 16 with adenocarcinomas, were examined for the expression of nm23-H1 by means of the immunohistochemical technique. RESULTS The total positive rate was 23.64% (13/55). The tissular nm23-H1 immunostaining had significantly higher expression in mucinous cancer (57.14%) than in other histologic types of epithelial ovarian cancer (P<0.05). Compared to that in the primary tumor, nm23-H1 expression in the omental metastatic tumor in the same patient was reduced. CONCLUSIONS The expression rate of nm23-H1 is higher in the primary tumors than in the metastatic tumors and is different in various histologic subtypes of epithelial ovarian cancer. The expression of nm23-H1 may participate in the inhibitory process of tumor invasion.

Methicillin resistant Staphyloccus aureus (MRSA) is a bacterium. The medical community requests development of new antibiotics to combat MRSA and many pharmaceutical companies are working to do this.

In China, the incidence of cleft lip and palate in almost the same as in other countries, ranging from 1.34‰ to 3.07‰ in new born babies. The population in China is the largest in the world, and the incidence of cleft lip and palate is also the largest.

Pregnancy induced hypertension (PIH) is a serious complication of pregnancy. Its basic pathophysiological change is well known as a generalized spasm of the smaller arterioles. In severe cases, if the cerebral vessels and involved, it could result in cerebral vessels are involved, it could result in cerebral ischemia, edema, infarction and haemorrhagia, and even cause death.

OBJECTIVE To investigate safety and De-qi (obtaining of needling sensation) depth of acupoint and their relation to therapeutic effect, and to electric resistance. METHODS 1) We plotted the graph to compare the differences of each acupoint depth between modern and ancient acupuncture writings; 2) 80 cadavers, and 240 subjects with computer tomography of chest to study the safety depth of acupoint were used, and their correlation to the length of the second phalanx of middle finger as Tong Shen Cun's standard; 3) 300 subjects were divided according to their body height and weight into normal-weight, over-weight and under-weight groups of 100 subjects to study the De-qi depth of acupoint; 4) using the 120 subjects which accepted acupuncture treatment due to pain symptom to study the relation between De-qi depth and therapeutic effect; 5) 107 subjects of different sizes were used to study the relation between De-qi depth and thickness of body, and electric resistance. RESULTS Acupoint depth was greater in modern acupuncture writings than in ancient writings. The safety depth of each acupoint in chest and in back was different, and they had high correlation to Tong Shen Cun's standard in adults, but not in newborns. The safety depths in chest acupoints were greater in female than in male, but not in back, and they related to body size. The De-qi depth was correlated with their therapeutic effects, corresponding to body thickness, but not related to their electric resistance. CONCLUSIONS Safety and De-qi depth of acupoint are related to body thickness. The length of the second phalanx of middle finger may be used as Tong Shen Cun's standard in adults, but not in newborns.